The human PCSK7 gene encodes proprotein convertase 1 (PC7), which is also known as PC8 and lymphoma PC (1, 2). As a serine protease of the furin/PC family, the deduced amino acid sequence of human PC7 consists of a signal peptide (residues 1 to 37), a propeptide (residue 38 to 141), and a mature chain (residues 142 to 785) that consists of extracellular (residues 142 to 667), transmembrane (residues 668 to 688) and cytoplasmic (residues 689 to 785) domains. Autocatalytic processing results in the removal of the propeptide, which confers potent inhibitory activity toward the enzyme (3-5). The purified recombinant human PCSK7 corresponds to the extracellular domain of the mature chain with a 10X His tag at the N-terminus. However, it may also contain the propeptide, as suggested by the N-terminal sequencing results. Thermolysin treatment increases its activity, which may be due to degradation of the propeptide by thermolysin.
Recombinant Human Proprotein Convertase 7/PCSK7 Protein, CF
R&D Systems | Catalog # 2984-SE
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Key Product Details
- R&D Systems NS0-derived Recombinant Human Proprotein Convertase 7/PCSK7 Protein (2984-SE)
- Quality control testing to verify active proteins with lot specific assays by in-house scientists
- All R&D Systems proteins are covered with a 100% guarantee
Source
NS0
Accession Number
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived human Proprotein Convertase 7/PCSK7 protein
Leu38-Thr667, with a 10-His tag between Arg141 and Ser142
Leu38-Thr667, with a 10-His tag between Arg141 and Ser142
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Leu38 & His tag
Predicted Molecular Mass
70 kDa (pro form) & 59 kDa (mature form)
SDS-PAGE
79 kDa, reducing conditions
Activity
Measured by its ability to cleave the fluorogenic peptide substrate pERTKR-AMC (Catalog # ES013).
The specific activity is >125 pmol/min/µg, as measured under the described conditions.
The specific activity is >125 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
2984-SE
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl, CaCl2 and Glycerol. |
| Shipping | The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Background: Proprotein Convertase 7/PCSK7
References
- Bruzzaniti, A. et al. (1996) Biochem. J. 314:727.
- Meerabux, J. et al. (1996) Cancer Res. 56:448.
- Van de Loo, J.-W.H.P. et al. (1997) J. Biol. Chem. 272:27116.
- Bhattacharjya, S. et al. (2000) Biochemistry 39:2868.
- Seidah, N.G. and M. Chretien (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, et al.) pp. 1877, Academic Press, San Diego.
Alternate Names
LPC, PCSK7, SPC7
Gene Symbol
PCSK7
UniProt
Additional Proprotein Convertase 7/PCSK7 Products
Product Documents for Recombinant Human Proprotein Convertase 7/PCSK7 Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human Proprotein Convertase 7/PCSK7 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human Proprotein Convertase 7/PCSK7 Protein, CF
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Protocols
View specific protocols for Recombinant Human Proprotein Convertase 7/PCSK7 Protein, CF (2984-SE):
Materials
- Activation Buffer: 50 mM Tris, 10 mM CaCl2, 0.15 M NaCl, 0.05% Brij 35, pH 7.5 (TCNB)
- Assay Buffer: 25 mM Tris, pH 7.0
- Recombinant Human Proprotein Convertase 7/PCSK7 (PCSK7) (Catalog # 2984-SE)
- Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
- 1,10 Phenanthroline (Sigma, Catalog # 320056), 0.6 M stock in DMSO
- Substrate: L-PyroGlu-Arg-Thr-Lys-Arg-AMC (Catalog # ES013)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescence Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Activate rhPCSK7 by diluting to 50 µg/mL in Activation Buffer containing 2.0 µg/mL Thermolysin.
- Incubate activation reaction at 37 °C for 4 hours.
- Add 1,10 Phenanthroline at a final concentration of 10 mM to stop Thermolysin activity.
- Dilute Substrate to 200 µM in Assay Buffer.
- Diluted activated rhPCSK7 to 2.0 µg/mL in Assay Buffer.
- In a plate load 50 µL of 2.0 µg/mL rhPCSK7 to wells and include a Substrate Blank of 50 µL of Assay Buffer.
- Start the reaction by adding 50 µL of 200 µM Substrate to wells.
- Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
- Calculate specific activity:
|
Specific Activity (pmol/min/µg) = |
Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU) |
| amount of enzyme (µg) |
*Adjusted for Substrate Blank
**Derived using calibration standard 7-amino, 4-Methyl Coumarin (Sigma, Catalog # A-9891).
Per Well:
- rhPCSK7: 0.1 µg
- Substrate: 100 µM
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